Annual Report 2022

Division of Cellular Signaling

Shinji Kohsaka, Toshihide Ueno, Yosuke Tanaka, Hiroyuki Mano

Introduction

 Through an approach combining high-sensitivity functional screening with next-generation sequencing analysis, we aim to elucidate the development mechanism of human tumors and develop novel molecular targeted therapies.

The Team and What We Do

• Identification of tumorigenesis and development of new drugs through comprehensive genome analysis.
• Establishment of functional assay to evaluate gene alterations.
• Development of new computational pipelines for cancer research.

Research Activities

1) We conducted comprehensive molecular profiling to discover novel predictive and prognostic biomarkers for salivary duct carcinoma (SDC). A total of 67 SDC samples were examined with DNA sequencing of 464 genes and transcriptome analysis in combination with the clinicopathological characteristics of the individuals. Prognostic biomarkers associated with response to combined androgen blockade (CAB) treatment were explored using mRNA expression data from 27 cases. Oncogenic mutations in receptor tyrosine kinase (RTK) genes or genes in the MAPK pathway were identified in 55 cases (82.1%). Alterations in the phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathway were identified in 38 cases (56.7%). Interestingly, patient prognosis could be predicted using mRNA expression profiles, but not genetic mutation profiles. The risk score generated from the expression data of a four-gene set that includes the ADAMTS1, DSC1, RNF39, and IGLL5 genes was a significant prognostic marker for overall survival in the cohort (HR = 5.99, 95% confidence interval (CI) = 2.73-13.1, p = 7.8 × 10-6). Another risk score constructed from the expression of CD3E and LDB3 was a strong prognostic marker for progression-free survival for CAB treatment (p = 0.03). Mutations in RTK genes, MAPK pathway genes, and PI3K/AKT pathway genes likely represent key mutations in SDC tumorigenesis. The gene expression profiles identified in this study may be useful for stratifying patients who are good candidates for CAB treatment and may benefit from additional systemic therapies (Kohsaka S. et al., 6, 1, 82, NPJ Precis Oncol. 2022).

2) Activating mutations in mitogen-activated protein kinase kinase 1 (MAP2K1) are involved in a variety of cancers and may be classified according to their RAF dependence. Sensitivity to combined BRAF and MEK treatments is associated with co-mutations of MAP2K1 and BRAF; however, the significance of less frequent MAP2K1 mutations is largely unknown. We evaluated the transforming potential and drug sensitivity of 100 MAP2K1 using individual assays and the mixed-all-nominated-in-one method. In addition, A375, a melanoma cell line harboring the BRAF V600E mutation, was used to evaluate the function of the MAP2K1 variants in combination with active RAF signaling. Among a total of 67 variants of unknown significance, 16 were evaluated as oncogenic or likely oncogenic. The drug sensitivity of the individual variants did not vary with respect to BRAF inhibitors, MEK inhibitors (MEKi), or their combination. Sensitivity to BRAF inhibitors was associated with the RAF dependency of the MAP2K1 variants, whereas resistance was higher in RAF-regulated or independent variants compared with RAF-dependent variants. Thus, the synergistic effect of BRAF and MEKis may be observed in RAF-regulated and RAF-dependent variants. MAP2K1 variants exhibit differential sensitivity to BRAF and MEKis, suggesting the importance of individual functional analysis for the selection of optimal treatments for each patient. This comprehensive evaluation reveals precise functional information and provides optimal combination treatment for individual MAP2K1 variants (Mizono S. et al., 22, 2, 227-239, Mol Cancer Ther, 2023).

3) We conducted an analysis of WES samples obtained from patients who had undergone hematopoietic stem cell transplantation for acute myeloid leukemia, subsequently developing secondary cancer. During our genome analysis of secondary cancers, we encountered a challenge related to the detection of somatic mutations. Specifically, SNPs present in both the donor and recipient genomes had the potential to generate false-positive mutation calls. To mitigate this issue, we developed a specialized pipeline. Furthermore, to evaluate the suitability of PARP inhibitors for treatment, it was essential to calculate the Homologous Recombination Deficiency (HRD) score. HRD scores were derived from WES and WGS bam data using established tools. However, because these scores were based on coverage data, disparities in SNPs between the donor and recipient genomes were erroneously interpreted as structural abnormalities unrelated to the disease. Consequently, this led to the calculation of excessively high HRD scores. To address this challenge, we devised a method to distinguish genomic structural abnormalities caused by the disease from false-positive structural anomalies resulting from SNP data. This approach allowed us to accurately calculate the HRD score for each case, providing valuable insights into our research.

Education

 One trainee from The University of Tokyo, one from Kyushu University, two from Juntendo University, one from Jikei University, and one from Yokohama City University.

Future Prospects

 Clinical genomics is now being implemented in clinical practice and sequencing technologies including whole genome sequencing, long read sequencing, and single cell analysis are advancing remarkably. We aim to contribute to the development and further improvement of clinical medicine through research and development of sequencing and screening technologies, and identification of useful biomarkers.

List of papers published in 2022

Journal

1. Mizuno S, Ikegami M, Koyama T, Sunami K, Ogata D, Kage H, Yanagaki M, Ikeuchi H, Ueno T, Tanikawa M, Oda K, Osuga Y, Mano H, Kohsaka S. High-Throughput Functional Evaluation of MAP2K1 Variants in Cancer. Molecular cancer therapeutics, 22:227-239, 2023

2. Okuma HS, Yoshida H, Kobayashi Y, Arakaki M, Mizoguchi C, Inagaki L, Voon PJ, Malik Bin Ismail A, Fen Soo Hoo H, Yusak S, Severino B Imasa M, Nguyen Huy T, Thai Anh T, Kohsaka S, Mano H, Yonemori K, Nakamura K, Yatabe Y. Molecular pathology quality control in Southeast Asia: Results of a multiregional quality assurance study from MASTER KEY Asia. Cancer science, 114:2664-2673, 2023

3. Kobayashi H, Makise N, Shinozaki-Ushiku A, Zhang L, Ishibashi Y, Ikegami M, Tsuda Y, Kohsaka S, Ushiku T, Oda K, Miyagawa K, Aburatani H, Mano H, Tanaka S. Dramatic response to entrectinib in a patient with malignant peripheral nerve sheath tumor harboring novel SNRNP70-NTRK3 fusion gene. Genes, chromosomes & cancer, 62:47-51, 2023

4. Kusakabe M, Taguchi A, Tanikawa M, Wagatsuma R, Yamazaki M, Tsuchimochi S, Toyohara Y, Kawata A, Baba S, Ueno T, Sone K, Mori-Uchino M, Ikemura M, Matsunaga H, Nagamatsu T, Wada-Hiraike O, Kawazu M, Ushiku T, Takeyama H, Oda K, Kawana K, Mano H, Osuga Y. Cells with stem-like properties are associated with the development of HPV18-positive cervical cancer. Cancer science, 114:885-895, 2023

5. Hayashi T, Kishi M, Takamochi K, Hosoya M, Kohsaka S, Kishikawa S, Ura A, Sano K, Sasahara N, Suehara Y, Takahashi F, Saito T, Suzuki K, Yao T. Expression of paired box 9 defines an aggressive subset of lung adenocarcinoma preferentially occurring in smokers. Histopathology, 82:672-683, 2023

6. Kage H, Shinozaki-Ushiku A, Ishigaki K, Sato Y, Tanabe M, Tanaka S, Tanikawa M, Watanabe K, Kato S, Akagi K, Uchino K, Mitani K, Takahashi S, Miura Y, Ikeda S, Kojima Y, Watanabe K, Mochizuki H, Yamaguchi H, Kawazoe Y, Kashiwabara K, Kohsaka S, Tatsuno K, Ushiku T, Ohe K, Yatomi Y, Seto Y, Aburatani H, Mano H, Miyagawa K, Oda K. Clinical utility of Todai OncoPanel in the setting of approved comprehensive cancer genomic profiling tests in Japan. Cancer science, 114:1710-1717, 2023

7. Chiba Y, Sudo K, Kojima Y, Okuma H, Kohsaka S, Machida R, Ichimura M, Anjo K, Kurishita K, Okita N, Nakamura K, Kinoshita I, Takahashi M, Matsubara J, Kusaba H, Yonemori K, Takahashi M. A multicenter investigator-initiated Phase 2 trial of E7090 in patients with advanced or recurrent solid tumor with fibroblast growth factor receptor (FGFR) gene alteration: FORTUNE trial. BMC cancer, 22:869, 2022

8. Kohno T, Kato M, Kohsaka S, Sudo T, Tamai I, Shiraishi Y, Okuma Y, Ogasawara D, Suzuki T, Yoshida T, Mano H. C-CAT: The National Datacenter for Cancer Genomic Medicine in Japan. Cancer discovery, 12:2509-2515, 2022

9. Kawakita D, Nagao T, Takahashi H, Kano S, Honma Y, Hirai H, Saigusa N, Akazawa K, Tani K, Ojiri H, Tsukahara K, Ozawa H, Okami K, Kondo T, Togashi T, Fushimi C, Shimura T, Shimizu A, Okamoto I, Okada T, Imanishi Y, Watanabe Y, Otsuka K, Sakai A, Ebisumoto K, Sato Y, Yamazaki K, Ueki Y, Hanazawa T, Saito Y, Ando M, Matsuki T, Nakaguro M, Sato Y, Urano M, Utsumi Y, Kohsaka S, Saotome T, Tada Y. Survival benefit of HER2-targeted or androgen deprivation therapy in salivary duct carcinoma. Therapeutic advances in medical oncology, 14:17588359221119538, 2022

10. Fukuhara S, Oshikawa-Kumade Y, Kogure Y, Shingaki S, Kariyazono H, Kikukawa Y, Koya J, Saito Y, Tabata M, Yoshifuji K, Mizuno K, Miyagi-Maeshima A, Matsushita H, Sugiyama M, Ogawa C, Inamoto Y, Fukuda T, Sugano M, Yamauchi N, Minami Y, Hirata M, Yoshida T, Kohno T, Kohsaka S, Mano H, Shiraishi Y, Ogawa S, Izutsu K, Kataoka K. Feasibility and clinical utility of comprehensive genomic profiling of hematological malignancies. Cancer science, 113:2763-2777, 2022

11. Imai M, Nakamura Y, Sunami K, Kage H, Komine K, Koyama T, Amano T, Ennishi D, Kanai M, Kenmotsu H, Maeda T, Morita S, Sakai D, Bando H, Makiyama A, Suzuki T, Hirata M, Kohsaka S, Tsuchihara K, Naito Y, Yoshino T. Expert panel consensus recommendations on the use of circulating tumor DNA assays for patients with advanced solid tumors. Cancer science, 113:3646-3656, 2022

12. Sunami K, Naito Y, Komine K, Amano T, Ennishi D, Imai M, Kage H, Kanai M, Kenmotsu H, Koyama T, Maeda T, Morita S, Sakai D, Kohsaka S, Tsuchihara K, Saigusa Y, Yoshino T. Chronological improvement in precision oncology implementation in Japan. Cancer science, 113:3995-4000, 2022

13. Naito Y, Sunami K, Kage H, Komine K, Amano T, Imai M, Koyama T, Ennishi D, Kanai M, Kenmotsu H, Maeda T, Morita S, Sakai D, Watanabe K, Shirota H, Kinoshita I, Yoshioka M, Mamesaya N, Ito M, Kohsaka S, Saigusa Y, Yamamoto K, Hirata M, Tsuchihara K, Yoshino T. Concordance Between Recommendations From Multidisciplinary Molecular Tumor Boards and Central Consensus for Cancer Treatment in Japan. JAMA network open, 5:e2245081, 2022

14. Nejo T, Takayanagi S, Tanaka S, Shinozaki-Ushiku A, Kohsaka S, Nagata K, Yokoyama M, Sora S, Ushiku T, Mukasa A, Aburatani H, Mano H, Saito N. Primary Intracranial Spindle Cell Sarcoma, DICER1-Mutant, with MDM2 Amplification Diagnosed on the Basis of Extensive Molecular Profiling. Clinical medicine insights. Case reports, 15:11795476221131189, 2022

15. Kohsaka S, Tada Y, Ando M, Nakaguro M, Shirai Y, Ueno T, Kojima S, Hirai H, Saigusa N, Kano S, Tsukahara K, Togashi T, Ozawa H, Kondo T, Okami K, Takahashi H, Kawakita D, Fushimi C, Suzuki T, Shimizu A, Okamoto I, Okada T, Sato Y, Imanishi Y, Watanabe Y, Sakai A, Ebisumoto K, Sato Y, Urano M, Honma Y, Yamazaki K, Ueki Y, Hanazawa T, Saito Y, Shimura T, Nagao T, Mano H. Identification of novel prognostic and predictive biomarkers in salivary duct carcinoma via comprehensive molecular profiling. NPJ precision oncology, 6:82, 2022

16. Morinaga T, Inozume T, Kawazu M, Ueda Y, Sax N, Yamashita K, Kawashima S, Nagasaki J, Ueno T, Lin J, Ohara Y, Kuwata T, Yukami H, Kawazoe A, Shitara K, Honobe-Tabuchi A, Ohnuma T, Kawamura T, Umeda Y, Kawahara Y, Nakamura Y, Kiniwa Y, Morita A, Ichihara E, Kiura K, Enokida T, Tahara M, Hasegawa Y, Mano H, Suzuki Y, Nishikawa H, Togashi Y. Mixed Response to Cancer Immunotherapy is Driven by Intratumor Heterogeneity and Differential Interlesion Immune Infiltration. Cancer research communications, 2:739-753, 2022

17. Kumagai S, Koyama S, Itahashi K, Tanegashima T, Lin YT, Togashi Y, Kamada T, Irie T, Okumura G, Kono H, Ito D, Fujii R, Watanabe S, Sai A, Fukuoka S, Sugiyama E, Watanabe G, Owari T, Nishinakamura H, Sugiyama D, Maeda Y, Kawazoe A, Yukami H, Chida K, Ohara Y, Yoshida T, Shinno Y, Takeyasu Y, Shirasawa M, Nakama K, Aokage K, Suzuki J, Ishii G, Kuwata T, Sakamoto N, Kawazu M, Ueno T, Mori T, Yamazaki N, Tsuboi M, Yatabe Y, Kinoshita T, Doi T, Shitara K, Mano H, Nishikawa H. Lactic acid promotes PD-1 expression in regulatory T cells in highly glycolytic tumor microenvironments. Cancer cell, 40:201-218.e9, 2022

18. Kawazu M, Ueno T, Saeki K, Sax N, Togashi Y, Kanaseki T, Chida K, Kishigami F, Sato K, Kojima S, Otsuka M, Kawazoe A, Nishinakamura H, Yuka M, Yamamoto Y, Yamashita K, Inoue S, Tanegashima T, Matsubara D, Tane K, Tanaka Y, Iinuma H, Hashiguchi Y, Hazama S, Khor SS, Tokunaga K, Tsuboi M, Niki T, Eto M, Shitara K, Torigoe T, Ishihara S, Aburatani H, Haeno H, Nishikawa H, Mano H. HLA Class I Analysis Provides Insight Into the Genetic and Epigenetic Background of Immune Evasion in Colorectal Cancer With High Microsatellite Instability. Gastroenterology, 162:799-812, 2022

19. Yasuda T, Sanada M, Kawazu M, Kojima S, Tsuzuki S, Ueno H, Iwamoto E, Iijima-Yamashita Y, Yamada T, Kanamori T, Nishimura R, Kuwatsuka Y, Takada S, Tanaka M, Ota S, Dobashi N, Yamazaki E, Hirose A, Murayama T, Sumi M, Sato S, Tange N, Nakamura Y, Katsuoka Y, Sakaida E, Kawamata T, Iida H, Shiraishi Y, Nannya Y, Ogawa S, Taniwaki M, Asou N, Hatta Y, Kiyoi H, Matsumura I, Horibe K, Mano H, Naoe T, Miyazaki Y, Hayakawa F. Two novel high-risk adult B-cell acute lymphoblastic leukemia subtypes with high expression of CDX2 and IDH1/2 mutations. Blood, 139:1850-1862, 2022

20. Jo H, Yagishita S, Hayashi Y, Ryu S, Suzuki M, Kohsaka S, Ueno T, Matsumoto Y, Horinouchi H, Ohe Y, Watanabe SI, Motoi N, Yatabe Y, Mano H, Takahashi K, Hamada A. Comparative Study on the Efficacy and Exposure of Molecular Target Agents in Non-small Cell Lung Cancer PDX Models with Driver Genetic Alterations. Molecular cancer therapeutics, 21:359-370, 2022

21. Nagasaki J, Inozume T, Sax N, Ariyasu R, Ishikawa M, Yamashita K, Kawazu M, Ueno T, Irie T, Tanji E, Morinaga T, Honobe A, Ohnuma T, Yoshino M, Iwata T, Kawase K, Sasaki K, Hanazawa T, Kochin V, Kawamura T, Matsue H, Hino M, Mano H, Suzuki Y, Nishikawa H, Togashi Y. PD-1 blockade therapy promotes infiltration of tumor-attacking exhausted T cell clonotypes. Cell reports, 38:110331, 2022

22. Ando T, Kage H, Shinozaki-Ushiku A, Tatsuno K, Tsutsumi S, Nagayama K, Nakajima J, Kohsaka S, Miyagawa K, Aburatani H, Mano H, Nagase T. Composite Clonal Analysis Reveals Transition of NSCLC Subtypes Through Accumulation of Gene Mutations: A Case Report. JTO clinical and research reports, 3:100277, 2022

23. Taguchi A, Rokutan H, Oda K, Tanikawa M, Tanimoto S, Sone K, Mori M, Tsuruga T, Kohsaka S, Tatsuno K, Shinozaki-Ushiku A, Miyagawa K, Mano H, Aburatani H, Ushiku T, Osuga Y. Genetic diagnosis of pseudomyxoma peritonei originating from mucinous borderline tumor inside an ovarian teratoma. BMC medical genomics, 15:51, 2022

24. Kishigami F, Tanaka Y, Yamamoto Y, Ueno T, Kojima S, Sato K, Inoue S, Sugaya S, Ishihara S, Mano H, Kawazu M. Exploration of predictive biomarkers for postoperative recurrence of stage II/III colorectal cancer using genomic sequencing. Cancer medicine, 11:3457-3470, 2022

25. Kage H, Kohsaka S, Tatsuno K, Ueno T, Ikegami M, Zokumasu K, Shinozaki-Ushiku A, Nagai S, Aburatani H, Mano H, Oda K. Tumor mutational burden measurement using comprehensive genomic profiling assay. Japanese journal of clinical oncology, 52:925-929, 2022

26. Takamochi K, Hara K, Hayashi T, Kohsaka S, Takahashi F, Suehara Y, Shimokawa M, Suzuki K. Clinical relevance of PD-L2 expression in surgically resected lung adenocarcinoma. Lung cancer (Amsterdam, Netherlands), 168:50-58, 2022

27. Nagai S, Nishihara H, Suzuki T, Nishio K, Taniguchi H, Tsuchihara K, Nakamura K, Takamatsu R, Ueno T, Aburatani H, Kohno T, Kohsaka S. Recommendations related to the analytical equivalence assessment of gene panel testing. Cancer science, 113:3282-3290, 2022

28. Tabata J, Nakaoku T, Araki M, Yoshino R, Kohsaka S, Otsuka A, Ikegami M, Ui A, Kanno SI, Miyoshi K, Matsumoto S, Sagae Y, Yasui A, Sekijima M, Mano H, Okuno Y, Okamoto A, Kohno T. Novel Calcium-Binding Ablating Mutations Induce Constitutive RET Activity and Drive Tumorigenesis. Cancer research, 82:3751-3762, 2022

29. Suehara Y, Kohsaka S, Hayashi T, Akaike K, Kurisaki-Arakawa A, Sato S, Kobayashi E, Mizuno S, Ueno T, Morii T, Okuma T, Kurihara T, Hasegawa N, Sano K, Sasa K, Okubo T, Kim Y, Mano H, Saito T. Identification of a Novel MAN1A1-ROS1 Fusion Gene Through mRNA-based Screening for Tyrosine Kinase Gene Aberrations in a Patient with Leiomyosarcoma. Clinical orthopaedics and related research, 479:838-852, 2021

30. Nakamura IT, Ikegami M, Hasegawa N, Hayashi T, Ueno T, Kawazu M, Yagishita S, Goto Y, Shinno Y, Kojima Y, Takamochi K, Takahashi F, Takahashi K, Mano H, Kohsaka S. Development of an optimal protocol for molecular profiling of tumor cells in pleural effusions at single-cell level. Cancer science, 112:2006-2019, 2021

31. Yagishita S, Kato K, Takahashi M, Imai T, Yatabe Y, Kuwata T, Suzuki M, Ochiai A, Ohtsu A, Shimada K, Nishida T, Hamada A, Mano H. Characterization of the large-scale Japanese patient-derived xenograft (J-PDX) library. Cancer science, 112:2454-2466, 2021

32. Suzuka J, Tsuda M, Wang L, Kohsaka S, Kishida K, Semba S, Sugino H, Aburatani S, Frauenlob M, Kurokawa T, Kojima S, Ueno T, Ohmiya Y, Mano H, Yasuda K, Gong JP, Tanaka S. Rapid reprogramming of tumour cells into cancer stem cells on double-network hydrogels. Nature biomedical engineering, 5:914-925, 2021

33. Matsuno Y, Hyodo M, Suzuki M, Tanaka Y, Horikoshi Y, Murakami Y, Torigoe H, Mano H, Tashiro S, Yoshioka KI. Replication-stress-associated DSBs induced by ionizing radiation risk genomic destabilization and associated clonal evolution. iScience, 24:102313, 2021

34. Chida K, Kawazoe A, Kawazu M, Suzuki T, Nakamura Y, Nakatsura T, Kuwata T, Ueno T, Kuboki Y, Kotani D, Kojima T, Taniguchi H, Mano H, Ikeda M, Shitara K, Endo I, Yoshino T. A Low Tumor Mutational Burden and PTEN Mutations Are Predictors of a Negative Response to PD-1 Blockade in MSI-H/dMMR Gastrointestinal Tumors. Clinical cancer research, 27:3714-3724, 2021

35. Nagano M, Kohsaka S, Hayashi T, Ueno T, Kojima S, Shinozaki-Ushiku A, Morita S, Tsuda M, Tanaka S, Shinohara T, Omori Y, Sugaya F, Kato H, Narita Y, Nakajima J, Suzuki K, Takamochi K, Mano H. Comprehensive molecular profiling of pulmonary pleomorphic carcinoma. NPJ precision oncology, 5:57, 2021

36. Nakamura IT, Kohsaka S, Ikegami M, Ikeuchi H, Ueno T, Li K, Beyett TS, Koyama T, Shimizu T, Yamamoto N, Takahashi F, Takahashi K, Eck MJ, Mano H. Comprehensive functional evaluation of variants of fibroblast growth factor receptor genes in cancer. NPJ precision oncology, 5:66, 2021

37. Hasegawa N, Kohsaka S, Kurokawa K, Shinno Y, Takeda Nakamura I, Ueno T, Kojima S, Kawazu M, Suehara Y, Ishijima M, Goto Y, Kojima Y, Yonemori K, Hayashi T, Saito T, Shukuya T, Takahashi F, Takahashi K, Mano H. Highly sensitive fusion detection using plasma cell-free RNA in non-small-cell lung cancers. Cancer science, 112:4393-4403, 2021

38. Inano T, Araki M, Morishita S, Imai M, Kihara Y, Okuda M, Yang Y, Ito M, Osaga S, Mano H, Edahiro Y, Ochiai T, Misawa K, Fukuda Y, Ando J, Komatsu N. Cell-autonomous megakaryopoiesis associated with polyclonal hematopoiesis in triple-negative essential thrombocythemia. Scientific reports, 11:17702, 2021

39. Kobayashi H, Makise N, Shinozaki-Ushiku A, Ishibashi Y, Ikegami M, Kohsaka S, Ushiku T, Oda K, Miyagawa K, Aburatani H, Mano H, Tanaka S. Scapular Angiomatoid Fibrous Histiocytoma with EWSR1-CREB1 Fusion in an Adult Patient. Case reports in orthopedics, 2021:9434222, 2021

40. Kawashima S, Inozume T, Kawazu M, Ueno T, Nagasaki J, Tanji E, Honobe A, Ohnuma T, Kawamura T, Umeda Y, Nakamura Y, Kawasaki T, Kiniwa Y, Yamasaki O, Fukushima S, Ikehara Y, Mano H, Suzuki Y, Nishikawa H, Matsue H, Togashi Y. TIGIT/CD155 axis mediates resistance to immunotherapy in patients with melanoma with the inflamed tumor microenvironment. Journal for immunotherapy of cancer, 9:e003134, 2021

41. Namba S, Ueno T, Kojima S, Kobayashi K, Kawase K, Tanaka Y, Inoue S, Kishigami F, Kawashima S, Maeda N, Ogawa T, Hazama S, Togashi Y, Ando M, Shiraishi Y, Mano H, Kawazu M. Transcript-targeted analysis reveals isoform alterations and double-hop fusions in breast cancer. Communications biology, 4:1320, 2021

42. Okuma HS, Yonemori K, Kojima Y, Tanioka M, Sudo K, Noguchi E, Hijioka S, Wakakuwa K, Kato K, Hirakawa A, Kuchiba A, Kubo T, Ichikawa H, Yoshida A, Yatabe Y, Nakamura K, Mano H, Yamamoto N, Fujiwara Y. Clinical Utility of Circulating Tumor DNA in Advanced Rare Cancers. Frontiers in oncology, 11:732525, 2021

43. Ikegami M, Kohsaka S, Hirose T, Ueno T, Inoue S, Kanomata N, Yamauchi H, Mori T, Sekine S, Inamoto Y, Yatabe Y, Kobayashi H, Tanaka S, Mano H. MicroSEC filters sequence errors for formalin-fixed and paraffin-embedded samples. Communications biology, 4:1396, 2021

44. Tanaka Y, Chiwaki F, Kojima S, Kawazu M, Komatsu M, Ueno T, Inoue S, Sekine S, Matsusaki K, Matsushita H, Boku N, Kanai Y, Yatabe Y, Sasaki H, Mano H. Multi-omic profiling of peritoneal metastases in gastric cancer identifies molecular subtypes and therapeutic vulnerabilities. Nature cancer, 2:962-977, 2021